AU772745B2

AU772745B2 - Substituted N-Benzyl-indol-3-YL glyoxylic acid derivatives having an anti-tumoral effect

Info

Publication number: AU772745B2
Application number: AU20119/01A
Authority: AU
Inventors: Gerald Bacher; Peter Emig; Eckhard Gunther; Guillaume Le Baut; Bernd Nickel; Dietmar Reichert
Original assignee: Zentaris AG
Current assignee: Aeterna Zentaris GmbH
Priority date: 1999-12-23
Filing date: 2000-12-19
Publication date: 2004-05-06
Anticipated expiration: 2020-12-19
Also published as: CA2395259A1; IL150235A0; US6432987B2; KR20020063245A; RU2266280C2; WO2001047913A2; MXPA02006229A; SK8752002A3; PL355684A1; DE50005284D1; DE19962300A1; BR0016712A; CN1283637C; CO5251472A1; RU2002120462A; JP2003519137A; AU2011901A; HUP0203716A2; CZ20022094A3; UA75060C2

Description

1 Substituted N-benzylindol-3-ylglyoxylic acid derivatives having antitumor action Indole-3-glyoxylamides have a variety of uses as pharmacodynamically active compounds and as synthetic building blocks in pharmaceutical chemistry.

In the patent application Neth. Appl. 6502481, compounds are described which have an antiinflammatory and antipyretic activity profile and analgesic activity.

In the British Application GB-B 1 028 812, derivatives of indolyl-3-glyoxylic acid and their amides are mentioned as analgesic, anticonvulsant and P-adrenergic compounds.

G. Domschke et al. (Ber. 94, 2353 (1961)) describes [sic] 3-indolylglyoxylamides which are not characterized pharmacologically.

E. Walton reports in J. Med. Chem., 11, 1252 (1968) on indolyl-3-giyoxylic acid derivatives which have an inhibitory action on glycerophosphate dehydrogenase and lactate dehydrogenase.

In the European Patent Specification EP 675110, 1H-indole-3-glyoxylamides are described which are profiled as sPLA2 inhibitors and are used in the treatment of septic shock, in pancreatitis and in the treatment of allergic rhinitis and rheumatoid arthritis.

It has already been proposed in the German Patent Application having the file reference 19814838.0 to employ the compounds according to DE-A 196 36 150 Al as antitumor agents.

2 The aim of the present is to make available novel compounds from the indol-3ylglyoxylic acid series of the general formula 1 as defined herein which have a good antitumor action and can be employed for the preparation of antiturior agents.

According to a first aspect of the invention there is provided a substituted Ns benzylindol-3-ylglyoxylic acid derivative having antitumor action of the general formula

I

Z R, R4

I

NNR2 z R3 N

-R

Formula 1 where the radicals R, RI, R 2

R

3

R

4 and Z have the following meaning: R is selected from nitro, amino, mono- or di(Ci-C 6 )-alkylamino, mono- or di(CI-Co)-cycloalkylamino, (CI-C6)-acylamino, phenyl (C -Cs)-alkylamino, aroylamino, hetero-aroylamino,

(C

1 -C6)-alkylsulfonamido, aryl-sulfonamido, maleimido, succinimido, phthalimido, benzyloxycarbonylamino, tert-butoxy-carbonylamino (BOC-amino), 9 -fluorenylmethoxy-carbonylamino (Fmoc-amino), triphenylmethylamino (Tr-amino), 2-(4 -pyridyl)ethoxycarbonylamino (Pyoc-amino), and diphenylmethylsilylamino (DPMS-amino), where the radicals for R can alternatively be substituted on the C atoms 2, 3 and 4 of the phenyl ring, S* R can furthermore be, in the case in which R 1 is hydrogen, methyl group, phenylmethyl group, the benzyloxycarbonyl radical, the tert-butoxycarbonyl radical (BOC radical) or the acetyl group, the following radicals: [l:\DayLib\LIBH]04581 .doc:UG

-NIJ-CH

2 -COOH; -NH-CH(CH 3 )-COOH; (CH3)2CH-CH 2

-CH

2

CH(NII>-COOH.

H3C-CH 2

-CH(CH

3 )-CH(NH)>COOH; HOH2C-CH(NI-)-COOH; phenyl-CH 2 CH(NI-)-COOH; (4-imidazoyl)-CH 2 -CH(NH~)COOH; H7N=C(NI{ 2

)-NII-(CH

2 3 ~CH(NHi)-COOH; H2N-(CH 2 4 -CH(NH)-COOH; H2N-CO-CH 2 -CH(NHj>COOH; or

HOOC-(CH

2 2

-CH(NIJ>COOH;

R, is selected from hydrogen, (CI-C6)-alkyl, where the alkyl group can be mono- or polysubstituted by a phenyl ring and this phenyl ring for its part can be mono- or polysubstituted by halogen, (CI-C6)-alkyl, (C3-C 7 )cycloalkyl, carboxyl groups, carboxyl o0 groups esterified with CI-C 6 -alkanols, trifluoromethyl groups, hydroxyl groups, methoxy groups, ethoxy groups, benzyloxy groups, a benzyl group which is mono- or polysubstituted in the phenyl moiety by (CI-C 6 )-alkyl groups, halogen atoms or trifluoromethyl groups, R, can further be a benzyloxycarbonyl group or a tertiary-butoxycarbonyl radical (Boc radical), or furthermore an acetyl group; R2can be a phenyl ring which is mono- or polysubstituted by (CI-C 6 )-alkyl, (C -CO)-alkoxy, cyano, halogen, trifluoromethyl, hydroxyl, benzyloxy, nitro, amino, -C6)-alkyalmino,

(C

1 -C6)-alkoxycarbonylamino, a carboxyl group, a carboxyl group esterified with C 1-C 6 -alkanols, or a pyridine structure of the formula 2 R54 20 6formula 2 or its N-oxide, where the pyridine structure is alternatively bonded to the ring carbon atoms 2, 3 and 4 and can be substituted by the substituents

R

5 and R 6 wherein R 5 and R 6 **fee*in formula 2 can be identical or different and are selected from (CI-C6)-alkyl,

(C

3

-C

7 cycloalkyl,

(C

1 -C6)-alkoxy, nitro, amino, hydroxyl, halogen, tri fluoromethyl, ethoxycarbonylamino radical, and carboxyalkyloxy in which the alkyl group can have 1-4 0 C atoms, *0 .0 00.00 OV 0 [1:\DayLib\LIBH]0458 1.doc:LJG 4 R2 can further be a 2- or 4-pyrimidinyl heterocycle, where the 2-pyrimidinyl ring can be mono- or polysubstituted by a methyl group, R2 can furthermore be a 7- or 8-quinolyl structure substituted by (Ci-C6)-alkyl, halogen, a nitro group, an amino group, or a (CI-C6)-alkylamino radical, or a 3- or 4-quinolylmethyl group, where the ring carbons of the pyridylmethyl radical of the quinolyl group and the quinolylmethyl radical can be substituted by (C-C 6 )-alkyl, (Ci-C 6 )-alkoxy, nitro, amino and (Ci-C6)-alkoxycarbonylamino, RI and Rz can further form, together with the nitrogen atom to which they are bonded, a piperazine ring of the formula 3 or a homopiperazine ring, provided R 2 is an aminoalkylene group, -N

N-R

7 Formula 3 in which R 7 is an alkyl radical, a phenyl ring which can be mono- or polysubstituted by (Ci-C 6 )-alkyl, (Ci-C 6 )-alkoxy, halogen, nitro, amino or (Ci-C6)-alkylamino;

R

7 can furthermore be a benzhydryl group or a bis-p-fluorobenzhydryl group; [R:\LIBH14773.doc aak

R

3 and R 4 can be identical or different and are individually selected from hydrogen,

(CI-C

6 )-alkyl, (C 3

-C

7 )cycloalkyl, (CI-C 6 )-alkanoyl, halogen and benzyloxy; R 3 and R 4 can furthermore be individually selected from a nitro group, an amino group, (C 1

-C

4 mono- or dialkyl-substituted amino group, (CI-C 6 )-alkoxycarbonylamino function, and 1 I-C 6 )-alkoxycarbonylamino-(CI -C 6 )-alkyl function; and Z is selected from 0 and S.

The designation alkyl, alkanol, alkoxy or alkylamino group for the radicals R, RI,

R

2

R

3

R

4

R

5

R

6

R

7 is normally understood as meaning both "straight-chain" [I:\DayLib\LIBH]O 4 5 8 I .doc:LIG 6 and "branched" alkyl groups, where "straight-chain alkyl groups" can be, for example, radicals such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl and "branched alkyl groups" designate, for example, radicals such as isopropyl or tert-butyl. "Cycloalkyl" is understood as meaning radicals such as, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. The designation "halogen" represents fluorine, chlorine, bromine or iodine. The designation "alkoxy group" represents radicals such as, for example, methoxy, ethoxy, propoxy, butoxy, isopropoxy, isobutoxy or pentoxy. The designation acyl of the acylamino radicals is to be understood as meaning the groups formyl, acetyl, propionyl, butyryl, valeryl and isovaleryl. The designation aroyl of the aroylamino groups represents benzoyl, naphthoyl, toluoyl, phthaloyl and the group heteroaroyl of the heteroaroylamino radicals represents nicotinoyl, isonicotinoyl, thenoyl and furoyl. The designation aryl of the arylsulfonamido group is understood as meaning phenyl, tolyl and naphthyl.

The compounds can aisc be employed as acid addition salts, for example as salts of minera- acids, such as, for example, hydrochloric acid, sulfuric acid, phosphoric acid, salts of organic acids, such as, for example, acetic acid, lactic acid, malonic acid, maleic acid, fumaric acid, gluconic acid, glucuronic acid, citric acid, ascorbic acid, embonic acid, methanesulfonic acid, trifluoroacetic acid, succinic acid, 2-hydroxyethanesulfonic acid, nicotinic acid and p-toluenesulfonic acid.

Both the compounds of the formula 1 and their salts are biologically active. The compounds of the formula 1 can be administered in free form or as salts with physiologically tolerable acids.

Administration can be performed orally, parenterally, intravenously, transdermally or by inhalation.

7 According to a second aspect of the invention there is provided the-compound

N-

(Pyridin-4-yl)-[ 1-( 4 -aminobenzyl)indol-3-yl]-glyoxylamide.

According to a third aspect of the invention there is provided the compound N- (Pyridin- 4 -yl)-[1-(4-nitrobenzyl)indol-3-yl]-glyoxylamide.

According to a fourth aspect of the invention there is provided acid addition salts of compounds of the general formula 1 according to any one of the first to third aspects of the invention as salts of mineral acids or salts of organic acids.

According to a fifth aspect of the invention there is provided pharmaceutical preparations comprising at least one compound of general formula 1 according to any one o0 of the first to third aspects of the invention, or a salt thereof according to the fourth aspect of the invention, together with physiologically tolerable inorganic or organic acids and, if appropriate, pharmaceutically utilisable vehicles and/or diluents or excipients.

Suitable administration forms are, for example, tablets, coated tablets, capsules, solutions for infusion or ampoules, suppositories, patches, powder preparations which can be employed by inhalation, suspensions, creams and ointments.

Accordingly, in a sixth aspect the invention provides application forms containing at least one compound of formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth aspect of the invention, in a form selected from tablets, coated tablets, capsules, solutions for infusion or ampoules, suppositories, patches, powder preparations which can be employed by inhalation, suspensions, creams and ointments.

According to a seventh aspect of the invention there is provided the use of at least one compound of general formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth aspect of the invention for the S* 25 manufacture of an antitumour composition.

According to an eighth aspect of the invention there is provided a method of treating tumours comprising administering to a subject in need of said treatment a therapeutically effective amount of at least one compound of formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth 30 aspect of the invention, or a pharmaceutical preparation according to the fifth aspect of the invention.

"According to a ninth aspect of the invention there is provided the use of at least one compound of general formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth aspect of the invention for-the treatment of tumours.

[I:\DayLib\LIBH]04581 .doc:UG 7a According to a tenth aspect of the invention there is provided the use"of at least one compound of general formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth aspect of the invention for the manufacture of a composition for treating oncoses.

According to an eleventh aspect of the invention there is provided a method of treating oncoses comprising administering to a subject in need of said treatment a therapeutically effective amount of at least one compound of formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth aspect of the invention, or a pharmaceutical preparation according to the fifth aspect of the invention.

According to a twelfth aspect of the invention there is provided the use of at least one compound of general formula 1 according to any one of the first to third aspects of the invention or a salt thereof according to the fourth aspect of the invention for the treatment of oncoses.

The processes for the preparation of the compounds according to the invention are described in the following reaction schemes 1 and 2 (Stages 1-3) and in general procedures. All compounds can be prepared as described or analogously.

The compounds of the general formula 1 with Z 0, R N0 2 and NH 2 and R 2 aryl, aralkyl and heteroaryl are obtainable according to the following Scheme 1: a e a e [I:\DayLib\LIBH]0458 .doc:UG 8 Scheme 1 ist stage: The indole derivative, which can be unsubstituted or monosubstituted or polysubstituted on C-2 or in the phenyl structure, is dissolved in a protic, dipolar aprotic or nonpolar organic solvent, such as, for example, isopropanol, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dioxane, toluene or methylene chloride and added dropwise to a suspension of a base prepared in a three-necked flask under an N 2 atmosphere or employed in a molar amount or in excess, such as, for example, sodium hydride, powdered potassium hydroxide, potassium tert-butoxide, dimethylaminopyridine or sodium amide, in a suitable solvent. Then the desired alkyl, aralkyl or heteroaralkyl halide, for 9 example, is added, if appropriate with addition of a catalyst, such as, for example, copper and the mixture is allowed to react for some time, for example for minutes to 12 hours, and the temperature is maintained within a range from 0°C to 120°C, preferably between 300C to [sic] 80°C, particularly between 500C and 650C.

After completion of the reaction, the reaction mixture is added to water, the solution is extracted, e.g. with diethyl ether, dichloromethane, chloroform, methyl tert-butyl ether or tetrahydrofuran, and the organic phase obtained in each case is dried with anhydrous sodium sulfate. The organic phase is concentrated in vacuo, the residue which remains is crystallized by trituration or the oily residue is purified by recrystallization, distillation or by column or flash chromatography on silica gel or alumina. The eluent used is, for example, a mixture of dichloromethane and diethyl ether in the ratio 8:2 (vol/vol) or a mixture of dichloromethane and ethanol in the ratio 9:1 (vol/vol).

2nd stage The N-substituted indole obtained accordino to the above procedure of the 1st stage is dissolved under a nitrogen atmosphere in an aprotic or nonpolar organic solvent, such as, for example, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, dioxane, toluene, xylene, methylene chloride or chloroform and added to a solution prepared under a nitrogen atmosphere of a monomolar up to 60% excess amount of oxalyl chloride in an aprotic or nonpolar solvent, such as, for example, in diethyl ether, methyl tert-butyl ether, tetrahydrofuran, dioxane, toluene, xylene, methylene chloride, the temperature being kept between -5°C and The reaction solution is then heated at a temperature between 10°C and 130°C, preferably between 200C and 800C, particularly between 30°C and 50°C, for a period of 30 minutes to 5 hours and the solvent is 10 then evaporated. The residue of the "indolyl- 3-glyoxyloyl chloride" formed in this manner which remains is dissolved in an aprotic solvent such as, for example, tetrahydrofuran, dioxane, diethyl ether, toluene or alternatively in a dipolar aprotic solvent, such as, for example, dimethylformamide, dimethylacetamide or dimethyl sulfoxide, cooled to a temperature between 10°C and -150C, preferably between and 0°C, and treated in the presence of an acid scavenger with a solution of the primary or secondary amine in a diluent. Possible diluents are the solvents used above for dissolving the indolyl-3-glyoxyloyl chloride. Acid scavengers used are triethylamine, pyridine, dimethylaminopyridine, basic ion exchanger, sodium carbonate, potassium carbonate, powdered potassium hydroxide and excess primary or secondary amine employed for the reaction. The reaction takes place at a temperature from 0°C to 1200C, preferably at 20-80°C, particularly between 40 0 C and 60 0 C. After a reaction time of 1-3 hours and standing at room temperature for 24 hours, the hydrochloride of the acid scavenger is filtered, the filtrate is concentrated in vacuc and the residue is recrystallizec from an organic solvent or purified by column chromatography on silica gel or alumina. The eluent used is, for example, a mixture of dichloromethane and ethanol (95:5, vol/vol) 3rd stage The N-nitrobenzyl-substituted "indoleglyoxylamide" obtained according to the above procedure (2nd stage) is dissolved in a protic solvent, such as, for example, methanol, ethanol, propanol, isopropanol or butanol or in a nonpolar solvent, such as, for example, tetrahydrofuran, dioxane or glycol dimethyl ether or in a dipolar aprotic solvent, such as, for example, dimethyl sulfoxide, dimethylformamide, dimethylacetamide or N-methylpyrrolidone and the solution is treated with a hydrogenation catalyst such as, for 11 example, Raney nickel, palladium/carbon or platinum under a nitrogen atmosphere and with stirring. Hydrogen is passed into the suspension with moderate shaking at a gas pressure of 1-15 bar, preferably 2-10 bar, particularly at 4-6 bar and the temperature is raised to about 20 0 -80 0 C, preferably 30 0 -60 0 C, particularly to 450-55°C. If appropriate, after about 1 hour a further amount of catalyst is added and the hydrogenation is continued. The hydrogenation was complete after a reaction time of 4-10 hours. The catalyst was filtered off under a nitrogen atmosphere, the solvent was concentrated to dryness in vacuo and the colorless to yellowish residue was dried in vacuo at 40 0

C.

Working Examples According to this general procedure for stages 1-3, on which synthesis scheme 1 is based, the following compounds were synthesized which are evident from the following tabulated list [sic] detailing the respective chemical name.

Example N-(Pyridin-4-yl)-[j-(4-aminobenzyl)indo-3-yl]glyoxy3amide (D-68838) 1st stage 1-(4-Nitrobenzyl)indole A mixture of 5.28 g of sodium hydride (0.22 mol, mineral oil suspension) in 200 ml of dimethyl sulfoxide is treated with a solution of 23.4 g (0.2.mol) of indole in 100 ml of dimethyl sulfoxide. It is heated at 65 0 C for 1 hour, then allowed to cool and 37.7 g (0.22 mol) of 4-nitrobenzyl chloride are then added dropwise. The solution is heated to 60 0 C, kept at room temperature for 14 hours and then poured into 700 ml of water with stirring. The mixture is extracted in 12 portions with a total of 300 ml of methylene chloride, the organic phase is dried using anhydrous sodium sulfate, filtered and the filtrate is concentrated in vacuo. The residue is purified on a silica gel column (silica gel 60, Merck AG, Darmstadt; eluent methylene chloride/ethanol 9:1, v/v).

Yield: 43.9 g (87% of theory) MS: m/e 253 (M+H) 2nd stage N-(Pyridin-4-yl)-[1-(4-nitrobenzyl)indol-3-yl]glyoxylamide (D-68836) A solution of 4.50 ml of oxalyl chloride in 50 ml of ether is treated dropwise with a solution of 10.09 g (0.04 mol) of l-(4-nitrobenzyl)indole in 50 ml of ether at OC [sic] and under a nitrogen atmosphere. The mixture is heated at reflux temperature for 2 hours and the solvent is then evaporated. 100 ml of retrahydrofuran are added tc the residue, it is cooled tc -5°C and a solution of of [sic 9.32 c (0.099 mol) of 4-aminopyridine in 400 ml of tetrahydrofuran is added dropwise. The mixture is heated to reflux for 3 hours and allowed to stand at room temperature overnight. The 4-aminopyridine hydrochloride is filtered off with suction, the precipitate is washed with tetrahydrofuran, the filtrate is concentrated in vacuo and the residue is recrystallized from ethyl acetate.

Yield: 13.5 g (84% of theory) MS: m/e 401 (M+H) 3rd stage 13 N-(Pyridin-4-yl)-[1-(4-aminobenzyl)indol-3-yl]glyoxylamide (D-68838) A mixture of 200 mg of Raney nickel in 50 ml of dioxane is treated with a suspension of 320 mg (0.8 mmol) of N-(pyridin-4-yl)-[1-(4-nitrobenzyl)indol-3-yl]glyoxylamide in a solvent mixture of 150 ml of dioxane and ml of isopropanol. Hydrogen is passed into this suspension with shaking at a gas pressure of 5 bar and the temperature is kept at 30-35 0 C. After about 3 hours, a further 400 mg of Raney nickel are added and the hydrogenation is continued at 35 0 C and 5 bar for a further 8 hours with vigorous shaking. The catalyst is filtered off under an N 2 atmosphere, the filtrate is concentrated to dryness in vacuo and the residue is dried in vacuo at 40 0

C.

Yield: 273 mg (92% of theory) MS: m/e '371 (M+H) Furthermore, the compounds of the general formula 1 with Z=C, R=NO: and NH: and aryl, aralkyl, heteroaryl, heteroaralkyl and the allylaminocarbonyl-2methylprop-i-yl group can also be synthesized according to the synthesis route of Scheme 2: 14 Scheme 2 1. (COCI), NH- 1 st stage

NH

NaH, DMSO N ON /CH -CI O NO 2 o 2nd stage 0

NH_/-

H

2 /Raney nickel /N I o 3rd stage

N

N H N-(Pyridin-4-yl)-[1-(4-aminobenzyl)indol-3-yl]glyoxlamide 1st stage N-(Pyridin-4-yl)-(indol-3-yl)glyoxylamide A solution of 10 g (85.3 mmol) of indole in 100 ml of ether is added dropwise at 0°C to a solution of 9 ml of oxalyl chloride in 100 ml of anhydrous ether. The mixture is kept under reflux for 3 hours. A suspension of 12 g (127.9 mmol) of 4-aminopyridine in 500 ml of tetrahydrofuran is then added dropwise at the reaction mixture is heated to reflux temperature with stirring for 3 hours and allowed to stand overnight at room temperature. It is filtered, the precipitate is treated with water and the dried compound is purified on a silica gel column (silica gel 60, Merck AG, 15 Darmstadt) using the eluent methylene chloride /ethanol (10:1, v/v).

Yield: 9.8 g (43.3% of theory) MS: m/e 266 (M+H) 2nd stage N-(Pyridin-4-yl)-[l-(4-nitrobeflzyl)indoli3-yllglyoxylamide (D-68836) The N- (pyridin-4-yl) -(indol-3-yl)glyoxylamide obtained according to the 1st Stage (Scheme 2) is reacted with 4-nitrobenzyl chloride according to the "benzylation procedure" (page 5) and the compound N-(pyridin-4-yl)- (4-nitrobenzyl)indol-3-yllglyoxylamide obtained is isolated.

Yield: 64% of theory MS: m/e 401 (M+H) K -(Pviridin-4-yi)-f]j-(4-arninobenzlZY)ifdo'1SY-v--ivoxy-J amide (D-68838) The N-(pyridin-4-yl)-[1-(4-flitrobeflzyl)ifdol3yl] glyoxylamide obtained according to the 2nd Stage (Scheme 2) is catalytically hydrogenated according to the "hydrogenation procedure" (page 7) and the compound N- (pyridin-4-yl) (4-aminobenzyl) indol-3-yl] glyoxylamide obtained is isolated.

Yield: 94% of theory MS: m/e 371 (M+H) 16 General procedure for the preparation of the compounds of the general formula 1 according to Scheme 2 1st stage: The indole derivative, which can be unsubstituted or substituted on C-2 or in the phenyl ring, dissolved in a solvent, as, for example, indicated above for oxalyl chloride, is added dropwise at a temperature between -5°C and +5°C to a solution prepared under a nitrogen atmosphere of a monomolar up to 60% excess amount of oxalyl chloride in an aprotic or nonpolar solvent, such as, for example, in diethyl ether, methyl tert-butyl ether, tetrahydrofuran, dioxane or alternatively dichloromethane. The reaction. solution is then heated for 1 to 5 hours to a temperature between 10 0 C and 1200C, preferably between 20 0 C and 800C, particularly between 30°C and 60°C, and the solvent is then evaporated. The residue of the (indol-3-yl)glyoxyloyl chloride which remains is dissolved or suspended in an aprotic solvent, such as, for example, tetrahydrofuran, dioxane, diethyl ether, toluene or alternatively in a dipolar aprotic solvent, such as, for example, dimethylformamide, dimethylacetamide or dimethyl sulfoxide, cooled to a temperature between -10°C and +100C, preferably to -50C to 0°C, and treated in the presence of an acid scavenger with a solution of the primary or secondary amine in a diluent. Possible diluents are the solvents used for dissolving the "indolyl-3-glyoxyloyl chloride". Acid scavengers used are triethylamine, pyridine, dimethylaminopyridine, basic ion exchanger, sodium carbonate, potassium carbonate, powdered potassium hydroxide and excess primary or secondary amine employed for the reaction.

The reaction takes place at a temperature from 0°C to 1200C, preferably at 20-800C, particularly between 400C and 600C. After a reaction time of 1-4 hours and standing at room temperature for 24 hours, the mixture is filtered, the precipitate is digested with water, 17 filtered off with suction and dried in vacuo. The desired compound is purified by recrystallization in an organic solvent or by column chromatography on silica gel or alumina. The eluent used is, for example, a mixture of dichloromethane and ethanol (10:1, vol/vol).

2nd Stage The "indol-3-ylglyoxylamide" obtained according to the above procedure of the 1st Stage is dissolved in a protic, dipoplar aprotic or nonpolar organic solvent, such as, for example, in isopropanol, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dioxane, toluene or methylene chloride and added dropwise to a suspension of a base prepared in a three-necked flask under an N 2 atmosphere or employed in a molar amount or in excess, such as, for example, sodium hydride, powdered potassium hydroxide, potassium tert-butoxide, dimethylaminopyridine or sodium amide, in a suitable solvent. The desired alkyl, aralkyl or heteroaralkyl halide is then added either undiluted or in a diluent, which was also used, for example, for dissolvinc the "indol-3-yiqlyoxyiamide", if appropriate with addition of a catalyst, such as, for example, copper and the mixture is allowed to react for some time, e.g. for minutes to 12 hours, and the temperature is kept within a range between 0°C and 120°C, preferably between 30°C and 80°C, particularly between 50 0 C and 70 0 C. After completion of the reaction, the reaction mixture is added to water, the solution is extracted, for example, with diethyl ether, dichloromethane, chloroform, methyl tert-butyl ether, tetrahydrofuran or n-butanol and the organic phase obtained in each case is dried using anhydrous sodium sulfate.

The organic phase is concentrated in vacuo, the residue which remains is crystallized by trituration or the oily residue is purified by distillation or by column or flash chromatography on silica gel or alumina. The 18 eluent used is, for example, a mixture of methylene chloride and diethyl ether in the ratio 8:2 (vol/vol) or a mixture of methylene chloride and ethanol in the ratio 9:1 3rd stage The N-nitrobenzyl-substituted "indoleglyoxylamide obtained according to the above procedure (2nd stage) is dissolved in a protic solvent, such as, for example, methanol, ethanol, propanol or butanol or in a nonpolar solvent, such as, for example, tetrahydrofuran, dioxane or glycol dimethyl ether' or in a dipolar aprotic solvent, such as, for example, dimethyl sulfoxide, dimethylformamide, dimethylacetamide or Nmethylpyrrolidone and the solution is treated with a hydrogenation catalyst such as, for example, Raney nickel, palladium/carbon or platinum under a nitrogen atmosphere and with stirring. Hydrogen is passed into the suspension with moderate shaking at a gas pressure of 1-15 bar, preferably 2-10 bar, particularly at 4-6 bar and the temperature is raised to about 20 0 -80 0

C,

preferably 30 0 -60 0 C, particularly tc 45 0 -55 0 C. If appropriate, after about 1 hour c further amount of catalyst is added and the hydrogenation is continued.

The hydrogenation was complete after a reaction time of 4-6 hours. The catalyst was filtered off under a nitrogen atmosphere, the solvent was concentrated to dryness and the colorless to yellowish residue was dried in vacuo at 40 0

C.

According to this general procedure for stages 1-3, on which the synthesis scheme 2 is based, the compounds D-68836 and D-68838 were synthesized, which have also already been prepared according to the synthesis procedure of reaction scheme 1.

It was possible to show the activity of the present compounds in a tubulin polymerization assay. In 19 particular, proof was provided that D-68838 inhibits the polymerization of tubulin and thus exerts a destabilizing effect on microtubuli or the mitotic spindles.

Claims

1. A substituted N-benzylindol-3-ylglyoxylic acid derivative having antitumor action of the general formula 1 Z R 1 R4 I NN N R2 R3 N -R Formula 1 where the radicals R, R 1 R 2 R 3 R4 and Z have the following meaning: R is selected from nitro, amino, mono- or di(Ci-C 6 )-alkylamino, mono- or di(CI-Cs)-cycloalkylamino, (CI-C 6 )-acylamino, phenyl (CI-C 6 )-alkylamino, aroylamino, hetero- aroylamino, (CI-C 6 )-alkylsulfonamido, aryl-sulfonamido, maleimido, succinimido, phthalimido, benzyloxycarbonylamino, tert-butoxy-carbonylamino (BOC-amino),

9-fluorenylmethoxy-carbonylamino (Fmoc-amino), triphenylmethylamino (Tr-amino), 2-(4 -pyridyl)ethoxycarbonylamino (Pyoc-amino), and diphenylmethylsilylamino (DPMS- amino), where the radicals for R can alternatively be substituted on the C atoms 2, 3 and 4 of the phenyl ring, Is R can furthermore be, in the case in which R, is hydrogen, methyl group, phenylmethyl group, the benzyloxycarbonyl radical, the tert-butoxycarbonyl radical (BOC radical) or the acetyl group, the following radicals: -NH-CH 2 -COOH; -NH-CH(CH 3 )-COOH; (CH 3 2 CH-CH 2 -CH 2 -CH(NH)-COOH; H 3 C-CH 2 -CH(CH 3 )-CH(NH)-COOH; HOH 2 C-CH(NH)-COOH; phenyl-CH 2 -CH(NH)- 0 20 COOH; (4-imidazoyl)-CH 2 -CH(NH)-COOH; HN=C(NH 2 )-NH-(CH 2 3 -CH(NH)-COOH; H 2 N-(CH 2 4 -CH(NH)-COOH; H 2 N-CO-CH 2 -CH(NH)-COOH; or qHOOC-(CH 2 2 -CH(NH)-COOH; R, is selected from hydrogen, (C,-C 6 )-alkyl, where the alkyl group can be mono- or polysubstituted by a phenyl ring and this phenyl ring for its part can be mono- or polysubstituted 0 025 by halogen, (CI-C 6 )-alkyl, (C 3 -C 7 )-cycloalkyl, carboxyl groups, carboxyl groups esterified with C 1 -C 6 -alkanols, trifluoromethyl groups, hydroxyl groups, methoxy groups, ethoxy groups, benzyloxy groups, a benzyl group which is mono- or polysubstituted in the phenyl moiety by (CI-C 6 )-alkyl groups, halogen atoms or trifluoromethyl groups, R can further be a benzyloxycarbonyl group or a tertiary-butoxycarbonyl radical (Boc 30 radical), or furthermore an acetyl group; 0 3 radical), or furthen-nore an acetyl group; 21 R 2 can be a phenyl ring which is mono- or polysubstituted by (Ci-C 6 )-alkyl, (Ci-C 6 )-alkoxy, cyano, halogen, trifluoromethyl, hydroxyl, benzyloxy, nitro, amino, (Ci-C 6 )-alkyalmino, (Ci-C6)-alkoxycarbonylamino, a carboxyl group, a carboxyl group esterified with Ci-C 6 -alkanols, or a pyridine structure of the formula 2 R 5 4 N 2 R 6 formula 2 or its N-oxide, where the pyridine structure is alternatively bonded to the ring carbon atoms 2, 3 and 4 and can be substituted by the substituents Rs and R 6 wherein R 5 and R 6 in formula 2 can be identical or different and are selected from (Ci-C 6 )-alkyl, (C 3 -C 7 cycloalkyl, (Ci-C 6 )-alkoxy, nitro, amino, hydroxyl, halogen, trifluoromethyl, ethoxycarbonylamino radical, and carboxyalkyloxy in which the alkyl group can have 1-4 C atoms, R2 can further be a 2- or 4-pyrimidinyl heterocycle, where the 2-pyrimidinyl ring can be mono- or polysubstituted by a methyl group, R2 can furthermore be a 7- or 8-quinolyl structure substituted by (Ci-C 6 )-alkyl, halogen, a nitro group, an amino group, or a (Ci-C 6 )-alkylamino radical, or R 2 is a 3- or 4-quinolylmethyl group, where the ring carbons of the pyridylmethyl radical of the quinolyl group and the quinolylmethyl radical can be substituted by (Ci-C 6 alkyl, (Ci-C 6 )-alkoxy, nitro, amino and (CI-C6)-alkoxycarbonylamino, R 1 and R 2 can further form, together with the nitrogen atom to which they are bonded, a piperazine ring of the formula 3 or a homopiperazine ring, provided R 2 is an aminoalkylene group, -N N-R 7 Formula 3 in which R 7 is an alkyl radical, a phenyl ring which can be mono- or polysubstituted by (Ci-C 6 )-alkyl, (Ci-C 6 )-alkoxy, halogen, nitro, amino or (C1-C 6 )-alkylamino; R 7 can furthermore be a benzhydryl group or a bis-p-fluorobenzhydryl group; R 3 and R 4 can be identical or different and are individually selected from hydrogen, (Ci-C 6 )-alkyl, (C3-C 7 )-cycloalkyl, (Ci-C 6 )-alkanoyl, halogen and benzyloxy; R 3 and R 4 can furthermore be individually selected from a nitro group, an amino group, p I p. p p pp PP p p.. 1 [R:\LIBH]04773.doc:aak 22 (Ci-C 4 )-mono- or dialkyl-substituted amino group, (Ci-C6)-alkoxycarbonylamino function, and (Ci-C 6 )-alkoxycarbonylamino-(C 1 -C 6 )-alkyl function; and Z is selected from O and S. 2. N-(Pyridin-4-yl)-[1-(4-aminobenzyl)indol-3-yl]-glyoxylamide. 3. N-(Pyridin-4-yl)-[l-(4-nitrobenzyl)indol-3-yl]-glyoxylamide. 4. A substituted N-benzylindol-3-ylglyoxylic acid derivative, substantially as hereinbefore described with reference to any one of the examples. Acid addition salts of compounds of the general formula 1 according to any one of claims 1 to 4 as salts of mineral acids or salts of organic acids. 0o 6. The acid addition salt of claim 5, wherein said mineral acid is selected from hydrochloric acid, sulfuric acid and phosphoric acid. 7. The acid addition salt of claim 5, wherein said organic acid is selected from acetic acid, lactic acid, malonic acid, maleic acid, fumaric acid, gluconic acid, glucuronic acid, citric acid, ascorbic acid, embonic acid, methanesulfonic acid, trifluoroacetic acid, succinic acid, 2-hydroxy-ethanesulfonic acid, nicotinic acid and p-toluene-sulfonic acid. 8. Pharmaceutical preparations comprising at least one compound of general formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7, together with physiologically tolerable inorganic or organic acids and, if appropriate, pharmaceutically utilisable vehicles and/or diluents or excipients. 20 9. Application forms containing at least one compound of formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7, in a form selected from tablets, coated tablets, capsules, solutions for infusion or ampoules, suppositories, patches, powder preparations which can be employed by inhalation, suspensions, creams and ointments.

10. Use of at least one compound of formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7, for the manufacture of an antitumour composition. .11. A method of treating tumours comprising administering to a subject in need of said treatment a therapeutically effective amount of at least one compound of formula 30 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims to 7, or a pharmaceutical preparation according to claim 8.

12. Use of at least one compound of formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7, for the treatment of tumours. [R:\LIBH]04773.doc:aak 23

13. Use of at least one compound of formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7, for the manufacture of a medicament for treating oncoses.

14. A method of treating oncoses comprising administering to a subject in need of said treatment a therapeutically effective amount of at least one compound of formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7, or a pharmaceutical preparation according to claim 8. Use of at least one compound of formula 1 according to any one of claims 1 to 4 or a salt thereof according to any one of claims 5 to 7 for the treatment of oncoses. Dated 27 February, 2004 Zentaris GmbH Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON 00... *0O S [R:\LIBH]04773.doc:aak